Table I. Restoration of cholesterol transport in npc1-deficient CHO cells by expression of yeast NCR1 .
| Percentage of corrected cells per GFP-positive cells
|
||||
|---|---|---|---|---|
| Partial
|
Complete
|
|||
| DNA | CT60 | NPC1-trap | CT60 | NPC1-trap |
| vector | 6.7 ± 2.1 | 3.0 ± 1.7 | 0.7 ± 0.5 | 2.3 ± 1.5 |
| NCR1 | 18.7 ± 4.5a | 17.3 ± 0.6a | 17.0 ± 2.6a | 15.3 ± 2.5a |
| NCR1-Cterm | 14.3 ± 2.5 | 10.7 ± 4.0 | 56.7 ± 2.1b | 61.3 ± 5.1b |
CHO cells were cotransfected with pEGFP and pCR3.1 (vector), pCR3.1-NCR1 (NCR1), or pCR3.1-NCR1-Cterm (NCR1-Cterm), incubated in the presence of low density lipoproteins, stained with filipin to detect unesterified cholesterol, and imaged for GFP. Partial restoration was defined as the reduction of intensity or the number of fluorescent vesicles compared with vector control. Cells lacking filipin fluorescence were classified as completely corrected. Means ± SEM from three independent experiments.
a
Significant differences from vector transfection, P < 0.01.
b
Significant differences from vector transfection, P < 0.001.